Results of sedimentation velocity analytical centrifugation analysis (SV-AUC) of N130 in the absence or presence of increasing concentrations of the rpL5 peptide.

Total concentration in mg/ml.

Sedimentation coefficient taken from the ordinate maximum of each peak in the best-fit distribution at 20 °C with percentage protein amount in parenthesis. Sedimentation coefficient (-value) is a measure of the size and shape of a protein in a solution with a specific density and viscosity at a specific temperature.

Standard sedimentation coefficient (-value) in water at 20 °C.

Molar mass values (MW) taken from the distribution that was transformed to the distribution. The theoretical molar mass is in parenthesis.

Best-fit weight-average frictional ratio values taken from the distribution.

Fit of the data in the 100-300 min sedimentation time frame, for the analysis of the lower MW rpL5:N130 complex intermediates. The MW cutoff of ~ 300 kDa corresponds to a tetramer of N130 pentamers.

Dynamic parameters for the N-terminus were determined using a global
value; thus, the only residue-specific motional parameter for residues within the N-terminus was
(see Materials and Methods).

The second transition in the FA curve (starting at ~2.5:1 rpL5:N130) corresponded to phase separation (
Figure 3a
). To gain insight into conformational changes associated with phase separation, we performed single-molecule Förster resonance energy transfer ( smFRET) experiments with a mutant of N130, N130
Q15C/S125C
, that could be dually labeled with Alexa Fluor594 680 at sites within the pentamer core (Q15C) and the A2 acidic tract (S125C), respectively. The FRET efficiency (E
FRET
) for this dye pair within the droplet phase (E
FRET
~0.15) was dramatically reduced in comparison with that observed in the absence of rpL5 (E
FRET
~0.85;
Figure 5
), consistent with theextension of the A2 tract from the N130 pentamer core due to interactions with rpL5 molecules upon phase separation (
Clearance New Arrival Left Hook Meshpaneled Stretch Sports Bra Black PE Nation Buy Cheap Low Shipping Fee Outlet Popular 2018 Newest Cheap Online Gbvgb2c1J
). In contrast to the monodisperse character of the SANS curves for samples of rpL5 and N130 at 0:1 and 1:1 (rpL5:N130) stoichiometry (
Sale Reliable Womens AfDiscretePyGt Maternity Nightie Mary White Clearance For Nice Supply gr21fde
), the curve for the 1:3 sample indicated periodic structural organization within the liquid-like droplets (
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, red curve). We interpreted these features in terms of inter-molecular correlation distances (55 Å, 77 Å, and 119 Å; see Analysis methods) due to organization of N130 and rpL5 induced by phase separation. Notably, nascent, higher order structural organization was evident in the SANS curve for a solution of rpL5 and N130 with 2:1 stoichiometry (
Figure 3b
, green curve). Assembly of rpL5 and N130 into higher order, soluble intermediates at and above the 2:1 stoichiometric ratio was also demonstrated using sedimentation velocity analytical ultracentrifugation (SV-AUC;
Figure 3—figure supplement 1
,
Table 3
). The appearance of high molecular weight species within the detection range (<1 MDa) was accompanied by a progressive decrease in the total mass detected, likely due to sedimentation of rpL5:N130 droplets in the sample cell during the dead-time of the experiment. Above the phase separation threshold (>3:1 rpL5:N130), resonances for residues within the N130 core broadened beyond detection in 2D
1
H-
15
N TROSY spectra but not those for residues within the A2 tract (
Figure 4a
). Chemical shift values indicated that these residues remained disordered.

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Tagged

July 20, 2016
Posted by
Chelsea Jones

Hungarian artist
László
Moholy-Nagy
was prolific during his lifetime, seeding a pedagogic spirit that asserted the potential for developing creativity to be ever-experimental. He spenthis life creating a new, modern artistic sensibility and teaching others to do the same.
Moholy-Nagy’s enduring influence is carried on through those who draw inspiration from the styles and methods he helped spearhead,
particularly in design and photography
. Read on to discover more about his life and achievements.

László
Moholy-Nagy’s enduring influence is carried on through those who draw inspiration from the styles and methods he helped spearhead,
particularly in design and photography

Moholy-Nagy began to produce his first drawings as a respite from war, eventuallysustaining an injury while serving in the Austro-Hungarian army in WWI. After he wasdischarged in 1918,he completed his law degree and began to study art more seriously and crystalizehis modern ideas, joining a group called MA that preached the revolutionary aspects of art.
He found inspiration in the Russian Constructivists’ use of art to enact social reform in a rapidly modernizing world, and was also influenced by Dadaism and Suprematism.

He found inspiration in the Russian Constructivists’ use of art to enact social reform in a rapidly modernizing world, and was also influenced by Dadaism and Suprematism.

Expandinginto mediums beyond paintings, he experimented beyondthe camerain his
photograms
and photomontage,
expanding the potential of photographyto see pastwhat the human eye does
. Hisphotograms were created by exposing light-sensitive paper directly to light, creating photographic images without using a camera.

expanding the potential of photographyto see pastwhat the human eye does

After relocatingto Berlin in 1919, hekicked off his teaching career.
Moholy-Nagy was appointed master of the
Staatliches Bauhaus art school in 1923 by founder Walter Gropius
, where he took on the school’s guiding design philosophy and expanded it over his five year period there. Bauhaus
incubated a style of design central to the Modernist aesthetic,centered on combining art and architecture in ways that harmonized form withfunction.
It was here that Moholy-Nagy best cultivated his avant-garde sensibility and belief that art and technology should be united.

This script allows the user to interact with Postgres in several ways.

It can simply start Postgres:

Or, it can be used to run Postgres and pass parameters to the server:

Lastly, it could also be used to start a totally different tool, such as Bash:

The
VOLUME
instruction should be used to expose any database storage area, configuration storage, or files/folders created by your docker container. You are strongly encouraged to use
VOLUME
for any mutable and/or user-serviceable parts of your image.

If a service can run without privileges, use
USER
to change to a non-root user. Start by creating the user and group in the
Dockerfile
with something like
RUN groupadd -r postgres useradd --no-log-init -r -g postgres postgres
.

Consider an explicit UID/GID

Users and groups in an image are assigned a non-deterministic UID/GID in that the “next” UID/GID is assigned regardless of image rebuilds. So, if it’s critical, you should assign an explicit UID/GID.

Due to an
unresolved bug
in the Go archive/tar package’s handling of sparse files, attempting to create a user with a significantly large UID inside a Docker container can lead to disk exhaustion because
/var/log/faillog
in the container layer is filled with NULL (\0) characters. A workaround is to pass the
--no-log-init
flag to useradd. The Debian/Ubuntu
adduser
wrapper does not support this flag.

Avoid installing or using
sudo
as it has unpredictable TTY and signal-forwarding behavior that can cause problems. If you absolutely need functionality similar to
sudo
, such as initializing the daemon as
root
but running it as non-
root
), consider using
“gosu”
.

Lastly, to reduce layers and complexity, avoid switching
USER
back and forth frequently.

For clarity and reliability, you should always use absolute paths for your
WORKDIR
. Also, you should use
WORKDIR
instead of proliferating instructions like
RUN cd … do-something
, which are hard to read, troubleshoot, and maintain.

An
ONBUILD
command executes after the current
Dockerfile
build completes.
ONBUILD
executes in any child image derived
FROM
the current image. Think of the
ONBUILD
command as an instruction the parent
Dockerfile
gives to the child
Dockerfile
.

A Docker build executes
ONBUILD
commands before any command in a child
Dockerfile
.

If done correctly, I could expect the response to look something like this:

Finally, I had to choose a database to store the profiles. I landed on the
Realtime Database
from Firebase, because it's a NoSQL database that uses JSON as its storage format. I installed
firebase
and
firebase-admin
on npm, then required them at the top of my `app.js` file alongside everything else.

To get writing to and reading from the database working, I had to authenticate Firebase using a specially generated "Service Account" private key. I generated the key in the Service Accounts tab of my Firebase settings, and dropped the corresponding code below the rest of my configurations.

I took a look at the sample response in the
GET users/show endpoint
documentation and I determined I’d need to store the user’s name, handle (referred to in the docs as
screen_name
), location, description, and profile image URL. I also wanted to save the profile color each user set on Twitter to use as a highlight color for buttons, links, and other accents, so I put the following code inside the client library convenience method I had saved in my `app.js` file.

To prevent situations where differences in capitalization would result in two instances of the same profile (like @julesforrest vs @JulesForrest), I set the handle variable to lowercase.

Then I noticed the profile images being returned were way too small to be useful on a desktop display. By tinkering with the image URL, I found a larger image path that worked better for my needs.

People often have URLs, @handles, #hashtags and email addresses in their profile descriptions, but unfortunately, the Twitter API was returning each description as a simple string. Luckily, I found a tool called
Autolinker
that would search each string and build out anchor tags in the appropriate places.

To get it working, I installed it via npm, then required it at the top of my app.js file:

Basic usage looked pretty straightforward and it came with a few options out of the box that could be passed as an object to the second parameter, like opening each link in a new window or adding a class to each anchor tag.

However, I wanted to add each user’s custom profile color (saved earlier from the API response) as an inline style on that user’s links, which required writing a custom replacement function. While custom replacement functions can handle some pretty complex configurations, I only tweaked the example from the documentation slightly to add the inline color styles and open each link in a new window. It’s important to note the social network for mentions and hashtags needs to be specified at the top of object parameter to link them properly, which wasn’t immediately clear from the docs.